US20160057957A1 - Pepper hybrid sv3301hw - Google Patents
Pepper hybrid sv3301hw Download PDFInfo
- Publication number
- US20160057957A1 US20160057957A1 US14/589,873 US201514589873A US2016057957A1 US 20160057957 A1 US20160057957 A1 US 20160057957A1 US 201514589873 A US201514589873 A US 201514589873A US 2016057957 A1 US2016057957 A1 US 2016057957A1
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- pepper
- hybrid
- seed
- sv3301hw
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
- A01H5/08—Fruits
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- A01G1/001—
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
- A01H6/82—Solanaceae, e.g. pepper, tobacco, potato, tomato or eggplant
- A01H6/822—Capsicum sp. [pepper]
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
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- C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
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- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
- C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
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Definitions
- the present invention relates to the field of plant breeding and, more specifically, to the development of pepper hybrid SV3301HW and the inbred pepper lines SHY-114-1002 and HHY-FD-1186.
- the goal of vegetable breeding is to combine various desirable traits in a single variety/hybrid.
- Such desirable traits may include any trait deemed beneficial by a grower and/or consumer, including greater yield, resistance to insects or disease, tolerance to environmental stress, and nutritional value.
- Breeding techniques take advantage of a plant's method of pollination. There are two general methods of pollination: a plant self-pollinates if pollen from one flower is transferred to the same or another flower of the same plant or plant variety. A plant cross-pollinates if pollen comes to it from a flower of a different plant variety.
- Plants that have been self-pollinated and selected for type over many generations become homozygous at almost all gene loci and produce a uniform population of true breeding progeny, a homozygous plant.
- a cross between two such homozygous plants of different genotypes produces a uniform population of hybrid plants that are heterozygous for many gene loci.
- a cross of two plants each heterozygous at a number of loci produces a population of hybrid plants that differ genetically and are not uniform. The resulting non-uniformity makes performance unpredictable.
- Pedigree breeding and recurrent selection are examples of breeding methods that have been used to develop inbred plants from breeding populations. Those breeding methods combine the genetic backgrounds from two or more plants or various other broad-based sources into breeding pools from which new lines and hybrids derived therefrom are developed by selfing and selection of desired phenotypes. The new lines and hybrids are evaluated to determine which of those have commercial potential.
- the present invention provides a pepper plant of the hybrid designated SV3301HW, the pepper line SHY-114-1002 or pepper line HHY-FD-1186. Also provided are pepper plants having all the physiological and morphological characteristics of such a plant. Parts of these pepper plants are also provided, for example, including pollen, an ovule, scion, a rootstock, a fruit, and a cell of the plant.
- a plant of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 comprising an added heritable trait comprising an added heritable trait.
- the heritable trait may comprise a genetic locus that is, for example, a dominant or recessive allele.
- a plant of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 is defined as comprising a single locus conversion.
- an added genetic locus confers one or more traits such as, for example, herbicide tolerance, insect resistance, disease resistance, and modified carbohydrate metabolism.
- the trait may be conferred by a naturally occurring gene introduced into the genome of a line by backcrossing, a natural or induced mutation, or a transgene introduced through genetic transformation techniques into the plant or a progenitor of any previous generation thereof.
- a genetic locus may comprise one or more genes integrated at a single chromosomal location.
- the invention also concerns the seed of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186.
- the pepper seed of the invention may be provided as an essentially homogeneous population of pepper seed of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186. Essentially homogeneous populations of seed are generally free from substantial numbers of other seed. Therefore, seed of hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 may be defined as forming at least about 97% of the total seed, including at least about 98%, 99% or more of the seed.
- the seed population may be separately grown to provide an essentially homogeneous population of pepper plants designated SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186.
- tissue culture of regenerable cells of a pepper plant of hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 is provided.
- the tissue culture will preferably be capable of regenerating pepper plants capable of expressing all of the physiological and morphological characteristics of the starting plant, and of regenerating plants having substantially the same genotype as the starting plant. Examples of some of the physiological and morphological characteristics of the hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 include those traits set forth in the tables herein.
- regenerable cells in such tissue cultures may be derived, for example, from embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistils, flowers, seed and stalks. Still further, the present invention provides pepper plants regenerated from a tissue culture of the invention, the plants having all the physiological and morphological characteristics of hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186.
- processes for producing pepper seeds, plants and fruit, which processes generally comprise crossing a first parent pepper plant with a second parent pepper plant, wherein at least one of the first or second parent pepper plants is a plant of pepper line SHY-114-1002 or pepper line HHY-FD-1186.
- These processes may be further exemplified as processes for preparing hybrid pepper seed or plants, wherein a first pepper plant is crossed with a second pepper plant of a different, distinct genotype to provide a hybrid that has, as one of its parents, a plant of pepper line SHY-114-1002 or pepper line HHY-FD-1186. In these processes, crossing will result in the production of seed. The seed production occurs regardless of whether the seed is collected or not.
- the first step in “crossing” comprises planting seeds of a first and second parent pepper plant, often in proximity so that pollination will occur for example, mediated by insect vectors. Alternatively, pollen can be transferred manually. Where the plant is self-pollinated, pollination may occur without the need for direct human intervention other than plant cultivation.
- a second step may comprise cultivating or growing the seeds of first and second parent pepper plants into plants that bear flowers.
- a third step may comprise preventing self-pollination of the plants, such as by emasculating the flowers (i.e., killing or removing the pollen).
- a fourth step for a hybrid cross may comprise cross-pollination between the first and second parent pepper plants. Yet another step comprises harvesting the seeds from at least one of the parent pepper plants. The harvested seed can be grown to produce a pepper plant or hybrid pepper plant.
- the present invention also provides the pepper seeds and plants produced by a process that comprises crossing a first parent pepper plant with a second parent pepper plant, wherein at least one of the first or second parent pepper plants is a plant of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186.
- pepper seed and plants produced by the process are first generation (F 1 ) hybrid pepper seed and plants produced by crossing a plant in accordance with the invention with another, distinct plant.
- the present invention further contemplates plant parts of such an F 1 hybrid pepper plant, and methods of use thereof. Therefore, certain exemplary embodiments of the invention provide an F 1 hybrid pepper plant and seed thereof.
- the present invention provides a method of producing a plant derived from hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186, the method comprising the steps of: (a) preparing a progeny plant derived from hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186, wherein said preparing comprises crossing a plant of the hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 with a second plant; and (b) crossing the progeny plant with itself or a second plant to produce a seed of a progeny plant of a subsequent generation.
- the method may additionally comprise: (c) growing a progeny plant of a subsequent generation from said seed of a progeny plant of a subsequent generation and crossing the progeny plant of a subsequent generation with itself or a second plant; and repeating the steps for an additional 3-10 generations to produce a plant derived from hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186.
- the plant derived from hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 may be an inbred line, and the aforementioned repeated crossing steps may be defined as comprising sufficient inbreeding to produce the inbred line.
- step (c) it may be desirable to select particular plants resulting from step (c) for continued crossing according to steps (b) and (c).
- plants having one or more desirable traits By selecting plants having one or more desirable traits, a plant derived from hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 is obtained which possesses some of the desirable traits of the line/hybrid as well as potentially other selected traits.
- the present invention provides a method of producing food or feed comprising: (a) obtaining a plant of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186, wherein the plant has been cultivated to maturity, and (b) collecting at least one pepper from the plant.
- the genetic complement of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 is provided.
- the phrase “genetic complement” is used to refer to the aggregate of nucleotide sequences, the expression of which sequences defines the phenotype of, in the present case, a pepper plant, or a cell or tissue of that plant.
- a genetic complement thus represents the genetic makeup of a cell, tissue or plant
- a hybrid genetic complement represents the genetic make up of a hybrid cell, tissue or plant.
- the invention thus provides pepper plant cells that have a genetic complement in accordance with the pepper plant cells disclosed herein, and seeds and plants containing such cells.
- Plant genetic complements may be assessed by genetic marker profiles, and by the expression of phenotypic traits that are characteristic of the expression of the genetic complement, e.g., isozyme typing profiles. It is understood that hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 could be identified by any of the many well known techniques such as, for example, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang
- the present invention provides hybrid genetic complements, as represented by pepper plant cells, tissues, plants, and seeds, formed by the combination of a haploid genetic complement of a pepper plant of the invention with a haploid genetic complement of a second pepper plant, preferably, another, distinct pepper plant.
- the present invention provides a pepper plant regenerated from a tissue culture that comprises a hybrid genetic complement of this invention.
- any method that “comprises,” “has” or “includes” one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps.
- any plant that “comprises,” “has” or “includes” one or more traits is not limited to possessing only those one or more traits and covers other unlisted traits.
- the invention provides methods and compositions relating to plants, seeds and derivatives of pepper hybrid SV3301HW, pepper line SHY-114-1002 and pepper line HHY-FD-1186.
- Hybrid SV3301HW is a hot Hungarian wax pepper with typical fruit type, fruit shape, smell, pungency, and flavor of a hot Hungarian wax. It has higher yield, 783 g/plant, compared to 713 g/plant for the standard hybrid Inferno. SV3301HW produces 17.3 fruits per plant while Inferno produces 15.3 fruits per plant. This hybrid is grown primarily in the U.S. for harvest as white fruits for processing and fresh market, with secondary use as a fresh pepper in Mexico. It exhibits excellent fresh and processed fruit quality due to its bright white color and moderate capsaicin content.
- hybrid SV3301HW average 18.6 cm long ⁇ 3.4 cm wide, with typical Hungarian wax shape and smell. Immature color is white (very pale yellow in some areas), and fruits mature to red. Fruit pungency of hybrid SV3301HW is moderate, around 640 Scoville Heat Units (fresh basis). Fruit weight of hybrid SV3301HW is 45.2 g while that of ‘Inferno’ is 46.7 g. Plant height averaged 70 cm, with a semi-spreading plant habit. It is ready to harvest at the white stage at approximately 75 days after transplanting, compared to 72 days for the standard ‘Inferno’. SV3301HW is well-adapted to the main hot pepper growing areas of the U.S. and Mexico, with excellent fresh and processed fruit quality.
- Hybrid SV3301HW is resistant to Tomato Spotted Wilt Virus (TSWV) and races 0-3 of Xanthomonas campestris vesacatoria (X3R), whereas Inferno is susceptible to both.
- TSWV Tomato Spotted Wilt Virus
- X3R Xanthomonas campestris vesacatoria
- hybrid SV3301HW The parents of hybrid SV3301HW are SHY-114-1002 and HHY-FD-1186.
- the parent lines are uniform and stable, as is a hybrid produced therefrom. A small percentage of variants can occur within commercially acceptable limits for almost any characteristic during the course of repeated multiplication. However no variants are expected.
- a plant having the physiological and morphological characteristics of pepper hybrid SV3301HW and the parent lines thereof.
- Leaf length of blade medium (Atol, Blondy, short Marconi, Merit, Anthea) width of blade medium (Albargia, narrow Balaton, Danubia, Marconi, Merit) width 64.3 mm 31.2 mm length 116.6 mm 108.7 mm petiole length 54.1 mm 39.0 mm color dark green dark green color (RHS Color Chart value) 147A 147A intensity of green color dark (Dolmy, Tinto) dark mature leaf shape ovate (Balico, Sonar) lanceolate leaf and stem pubescence absent light undulation of margin medium (Tenor) very weak blistering very weak very weak profile in cross section moderately concave strongly concave (Doux italien, Favolor) glossiness weak (De Cayenne, medium Doux NYC long des Favolor) 5.
- Flower peduncle attitude semi-drooping (Blondy) erect flowers per leaf axil 1.2 1 calyx lobes 6.6 6.4 petals 6.9 6.3 diameter 29.9 mm 28.9 mm corolla color white white corolla throat markings yellow yellow anther color purple purple style length exceeds stamen exceeds stamen self-incompatibility present present 6.
- Plant habit compact compact attitude upright/erect upright/erect plant height 65.9 cm 64.9 cm plant width 60.5 cm 52.3 cm length of stem from cotyledon 15.0 cm 21.1 cm to first flower length of the third internode 80.0 mm 118.0 mm (from soil surface) length of stem medium long shortened internode (in upper present absent part) number of internodes between one to three the first flower and the shortened internodes stem: hairiness of nodes weak weak height tall tall basal branches few (2-3) none branch flexibility willowy rigid stem strength (breakage intermediate weak resistance) 4.
- Flower peduncle attitude semi-drooping semi-drooping flowers per leaf axil 1 1 calyx lobes 5.5 5.9 petals 5.6 6 diameter 20.2 mm 22.4 mm corolla color white white fruit diameter at calyx 33.2 mm 36.1 mm attachment fruit diameter at mid-point 32.2 mm 30.7 mm flesh thickness at mid-point 2.7 mm 2.9 mm average number of fruits per 22.7 25.3 plant % large fruits 30.50% weight range 50 4.50% weight range to 90 50 to 58 % medium fruits 39.90% weight range 30 38.40% weight range to 49 30 to 49 % small fruits 29.60% weight range 1 57.10% weight range to 29 1 to 29 average fruit weight 64.8 gm 41.6 gm fruit shape (longitudinal narrowly triangular horn-shaped section) fruit shape (cross section, at angular/triangular angular/triangular level of placenta) sinuation of pericarp at basal weak absent or very weak part sinuation of
- One aspect of the current invention concerns methods for producing seed of pepper hybrid SV3301HW involving crossing pepper lines SHY-114-1002 and HHY-FD-1186.
- hybrid SV3301HW, line SHY-114-1002, or line HHY-FD-1186 may be crossed with itself or with any second plant.
- Such methods can be used for propagation of hybrid SV3301HW and/or the pepper lines SHY-114-1002 and HHY-FD-1186, or can be used to produce plants that are derived from hybrid SV3301HW and/or the pepper lines SHY-114-1002 and HHY-FD-1186.
- Plants derived from hybrid SV3301HW and/or the pepper lines SHY-114-1002 and HHY-FD-1186 may be used, in certain embodiments, for the development of new pepper varieties.
- novel varieties may be created by crossing hybrid SV3301HW followed by multiple generations of breeding according to such well known methods.
- New varieties may be created by crossing with any second plant. In selecting such a second plant to cross for the purpose of developing novel lines, it may be desired to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination.
- inbreeding and selection take place to produce new varieties. For development of a uniform line, often five or more generations of selfing and selection are involved.
- Uniform lines of new varieties may also be developed by way of double-haploids. This technique allows the creation of true breeding lines without the need for multiple generations of selfing and selection. In this manner true breeding lines can be produced in as little as one generation.
- Haploid embryos may be produced from microspores, pollen, anther cultures, or ovary cultures. The haploid embryos may then be doubled autonomously, or by chemical treatments (e.g. colchicine treatment). Alternatively, haploid embryos may be grown into haploid plants and treated to induce chromosome doubling. In either case, fertile homozygous plants are obtained.
- any of such techniques may be used in connection with a plant of the invention and progeny thereof to achieve a homozygous line.
- Backcrossing can also be used to improve an inbred plant.
- Backcrossing transfers a specific desirable trait from one inbred or non-inbred source to an inbred that lacks that trait. This can be accomplished, for example, by first crossing a superior inbred (A) (recurrent parent) to a donor inbred (non-recurrent parent), which carries the appropriate locus or loci for the trait in question. The progeny of this cross are then mated back to the superior recurrent parent (A) followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent. After five or more backcross generations with selection for the desired trait, the progeny have the characteristic being transferred, but are like the superior parent for most or almost all other loci. The last backcross generation would be selfed to give pure breeding progeny for the trait being transferred.
- the plants of the present invention are particularly well suited for the development of new lines based on the elite nature of the genetic background of the plants.
- a second plant to cross with SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 for the purpose of developing novel pepper lines it will typically be preferred to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination.
- desirable traits may include, in specific embodiments, high seed yield, high seed germination, seedling vigor, high fruit yield, disease tolerance or resistance, and adaptability for soil and climate conditions.
- Consumer-driven traits, such as a fruit shape, color, texture, and taste are other examples of traits that may be incorporated into new lines of pepper plants developed by this invention.
- hybrid SV3301HW exhibits desirable traits, as conferred by pepper lines SHY-114-1002 and HHY-FD-1186.
- the performance characteristics of hybrid SV3301HW and pepper lines SHY-114-1002 and HHY-FD-1186 were the subject of an objective analysis of the performance traits relative to other varieties. The results of the analysis are presented in Table 2.
- plants described herein are provided modified to include at least a first desired heritable trait.
- Such plants may, in one embodiment, be developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to a genetic locus transferred into the plant via the backcrossing technique.
- the term single locus converted plant as used herein refers to those pepper plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to the single locus transferred into the variety via the backcrossing technique.
- backcrossing essentially all of the morphological and physiological characteristics, it is meant that the characteristics of a plant are recovered that are otherwise present when compared in the same environment, other than an occasional variant trait that might arise during backcrossing or direct introduction of a transgene.
- Backcrossing methods can be used with the present invention to improve or introduce a characteristic into the present variety.
- the parental pepper plant which contributes the locus for the desired characteristic is termed the nonrecurrent or donor parent. This terminology refers to the fact that the nonrecurrent parent is used one time in the backcross protocol and therefore does not recur.
- the parental pepper plant to which the locus or loci from the nonrecurrent parent are transferred is known as the recurrent parent as it is used for several rounds in the backcrossing protocol.
- recurrent parent the original variety of interest (recurrent parent) is crossed to a second variety (nonrecurrent parent) that carries the single locus of interest to be transferred.
- nonrecurrent parent the second variety that carries the single locus of interest to be transferred.
- the resulting progeny from this cross are then crossed again to the recurrent parent and the process is repeated until a pepper plant is obtained wherein essentially all of the morphological and physiological characteristics of the recurrent parent are recovered in the converted plant, in addition to the single transferred locus from the nonrecurrent parent.
- a suitable recurrent parent is an important step for a successful backcrossing procedure.
- the goal of a backcross protocol is to alter or substitute a single trait or characteristic in the original variety.
- a single locus of the recurrent variety is modified or substituted with the desired locus from the nonrecurrent parent, while retaining essentially all of the rest of the desired genetic, and therefore the desired physiological and morphological constitution of the original variety.
- the choice of the particular nonrecurrent parent will depend on the purpose of the backcross; one of the major purposes is to add some commercially desirable trait to the plant.
- the exact backcrossing protocol will depend on the characteristic or trait being altered and the genetic distance between the recurrent and nonrecurrent parents.
- progeny pepper plants of a backcross in which a plant described herein is the recurrent parent comprise (i) the desired trait from the non-recurrent parent and (ii) all of the physiological and morphological characteristics of pepper the recurrent parent as determined at the 5% significance level when grown in the same environmental conditions.
- New varieties can also be developed from more than two parents.
- the technique known as modified backcrossing, uses different recurrent parents during the backcrossing. Modified backcrossing may be used to replace the original recurrent parent with a variety having certain more desirable characteristics or multiple parents may be used to obtain different desirable characteristics from each.
- one, two, three or four genomic loci may be integrated into an elite line via this methodology.
- this elite line containing the additional loci is further crossed with another parental elite line to produce hybrid offspring, it is possible to then incorporate at least eight separate additional loci into the hybrid.
- additional loci may confer, for example, such traits as a disease resistance or a fruit quality trait.
- each locus may confer a separate trait.
- loci may need to be homozygous and exist in each parent line to confer a trait in the hybrid.
- multiple loci may be combined to confer a single robust phenotype of a desired trait.
- Single locus traits have been identified that are not regularly selected for in the development of a new inbred but that can be improved by backcrossing techniques.
- Single locus traits may or may not be transgenic; examples of these traits include, but are not limited to, herbicide resistance, resistance to bacterial, fungal, or viral disease, insect resistance, modified fatty acid or carbohydrate metabolism, and altered nutritional quality. These comprise genes generally inherited through the nucleus.
- Direct selection may be applied where the single locus acts as a dominant trait.
- the progeny of the initial cross are assayed for viral resistance and/or the presence of the corresponding gene prior to the backcrossing. Selection eliminates any plants that do not have the desired gene and resistance trait, and only those plants that have the trait are used in the subsequent backcross. This process is then repeated for all additional backcross generations.
- Selection of pepper plants for breeding is not necessarily dependent on the phenotype of a plant and instead can be based on genetic investigations. For example, one can utilize a suitable genetic marker which is closely genetically linked to a trait of interest. One of these markers can be used to identify the presence or absence of a trait in the offspring of a particular cross, and can be used in selection of progeny for continued breeding. This technique is commonly referred to as marker assisted selection. Any other type of genetic marker or other assay which is able to identify the relative presence or absence of a trait of interest in a plant can also be useful for breeding purposes. Procedures for marker assisted selection are well known in the art.
- Types of genetic markers which could be used in accordance with the invention include, but are not necessarily limited to, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).
- SSLPs Simple Sequence Length Polymorphisms
- RAPDs Randomly Amplified Polymorphic DNAs
- DAF Sequence Characterized Amplified Regions
- Many useful traits that can be introduced by backcrossing, as well as directly into a plant are those which are introduced by genetic transformation techniques. Genetic transformation may therefore be used to insert a selected transgene into a plant of the invention or may, alternatively, be used for the preparation of transgenes which can be introduced by backcrossing. Methods for the transformation of plants that are well known to those of skill in the art and applicable to many crop species include, but are not limited to, electroporation, microprojectile bombardment, Agrobacterium -mediated transformation and direct DNA uptake by protoplasts.
- friable tissues such as a suspension culture of cells or embryogenic callus or alternatively one may transform immature embryos or other organized tissue directly.
- pectolyases pectolyases
- microprojectile bombardment An efficient method for delivering transforming DNA segments to plant cells is microprojectile bombardment.
- particles are coated with nucleic acids and delivered into cells by a propelling force.
- Exemplary particles include those comprised of tungsten, platinum, and preferably, gold.
- cells in suspension are concentrated on filters or solid culture medium.
- immature embryos or other target cells may be arranged on solid culture medium.
- the cells to be bombarded are positioned at an appropriate distance below the macroprojectile stopping plate.
- An illustrative embodiment of a method for delivering DNA into plant cells by acceleration is the Biolistics Particle Delivery System, which can be used to propel particles coated with DNA or cells through a screen, such as a stainless steel or Nytex screen, onto a surface covered with target cells.
- a screen such as a stainless steel or Nytex screen
- the screen disperses the particles so that they are not delivered to the recipient cells in large aggregates.
- Microprojectile bombardment techniques are widely applicable, and may be used to transform virtually any plant species.
- Agrobacterium -mediated transfer is another widely applicable system for introducing gene loci into plant cells.
- An advantage of the technique is that DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast.
- Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium , allowing for convenient manipulations (Klee et al., Bio - Technology, 3(7):637-642, 1985).
- recent technological advances in vectors for Agrobacterium -mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate the construction of vectors capable of expressing various polypeptide coding genes.
- the vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes.
- Agrobacterium containing both armed and disarmed Ti genes can be used for transformation.
- Agrobacterium -mediated transformation In those plant strains where Agrobacterium -mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene locus transfer.
- the use of Agrobacterium -mediated plant integrating vectors to introduce DNA into plant cells is well known in the art (Fraley et al., Bio/Technology, 3:629-635, 1985; U.S. Pat. No. 5,563,055).
- Transformation of plant protoplasts also can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation, and combinations of these treatments (see, e.g., Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al., Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature, 312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986; Marcotte et al., Nature, 335:454, 1988). Transformation of plants and expression of foreign genetic elements is exemplified in Choi et al. ( Plant Cell Rep., 13: 344-348, 1994), and Ellul et al. ( Theor. Appl. Genet., 107:462-469, 2003).
- a number of promoters have utility for plant gene expression for any gene of interest including but not limited to selectable markers, scoreable markers, genes for pest tolerance, disease resistance, nutritional enhancements and any other gene of agronomic interest.
- constitutive promoters useful for plant gene expression include, but are not limited to, the cauliflower mosaic virus (CaMV) P-35S promoter, which confers constitutive, high-level expression in most plant tissues (see, e.g., Odel et al., Nature, 313:810, 1985), including in monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591, 1990; Terada and Shimamoto, Mol. Gen.
- CaMV cauliflower mosaic virus
- P-eFMV FMV promoter
- a variety of plant gene promoters that are regulated in response to environmental, hormonal, chemical, and/or developmental signals can also be used for expression of an operably linked gene in plant cells, including promoters regulated by (1) heat (Callis et al., Plant Physiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter, Schaffner and Sheen, Plant Cell, 3:997, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., EMBO J., 4:2723, 1985), (3) hormones, such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989), (4) wounding (e.g., wunl, Siebertz et al., Plant Cell, 1:961, 1989); or (5) chemicals such as methyl jasmonate
- organ-specific promoters e.g., Roshal et al., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988; Bustos et al., Plant Cell, 1:839, 1989.
- Exemplary nucleic acids which may be introduced to plants of this invention include, for example, DNA sequences or genes from another species, or even genes or sequences which originate with or are present in the same species, but are incorporated into recipient cells by genetic engineering methods rather than classical reproduction or breeding techniques.
- exogenous is also intended to refer to genes that are not normally present in the cell being transformed, or perhaps simply not present in the form, structure, etc., as found in the transforming DNA segment or gene, or genes which are normally present and that one desires to express in a manner that differs from the natural expression pattern, e.g., to over-express.
- exogenous gene or DNA is intended to refer to any gene or DNA segment that is introduced into a recipient cell, regardless of whether a similar gene may already be present in such a cell.
- the type of DNA included in the exogenous DNA can include DNA which is already present in the plant cell, DNA from another plant, DNA from a different organism, or a DNA generated externally, such as a DNA sequence containing an antisense message of a gene, or a DNA sequence encoding a synthetic or modified version of a gene.
- Non-limiting examples of particular genes and corresponding phenotypes one may choose to introduce into a pepper plant include one or more genes for insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pest tolerance such as genes for fungal disease control, herbicide tolerance such as genes conferring glyphosate tolerance, and genes for quality improvements such as yield, nutritional enhancements, environmental or stress tolerances, or any desirable changes in plant physiology, growth, development, morphology or plant product(s).
- insect tolerance such as a Bacillus thuringiensis (B.t.) gene
- pest tolerance such as genes for fungal disease control
- herbicide tolerance such as genes conferring glyphosate tolerance
- quality improvements such as yield, nutritional enhancements, environmental or stress tolerances, or any desirable changes in plant physiology, growth, development, morphology or plant product(s).
- structural genes would include any gene that confers insect tolerance including but not limited to a Bacillus insect control protein gene as described in WO 99/31248, herein incorporated by reference in its entirety, U.S. Pat. No. 5,689,052, herein incorporated by reference in its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by reference in their entirety.
- the structural gene can confer tolerance to the herbicide glyphosate as conferred by genes including, but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, herein incorporated by reference in its entirety, or glyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporated by reference in its entirety.
- the DNA coding sequences can affect these phenotypes by encoding a non-translatable RNA molecule that causes the targeted inhibition of expression of an endogenous gene, for example via antisense- or cosuppression-mediated mechanisms (see, for example, Bird et al., Biotech. Gen. Engin. Rev., 9:207, 1991).
- the RNA could also be a catalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desired endogenous mRNA product (see for example, Gibson and Shillito, Mol. Biotech., 7:125, 1997).
- a catalytic RNA molecule i.e., a ribozyme
- any gene which produces a protein or mRNA which expresses a phenotype or morphology change of interest is useful for the practice of the present invention.
- Allele Any of one or more alternative forms of a gene locus, all of which alleles relate to one trait or characteristic. In a diploid cell or organism, the two alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes.
- Backcrossing A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (F 1 ), back to one of the parents of the hybrid progeny. Backcrossing can be used to introduce one or more single locus conversions from one genetic background into another.
- F 1 first generation hybrid
- Crossing The mating of two parent plants.
- Cross-pollination Fertilization by the union of two gametes from different plants.
- Diploid A cell or organism having two sets of chromosomes.
- Emasculate The removal of plant male sex organs or the inactivation of the organs with a cytoplasmic or nuclear genetic factor or a chemical agent conferring male sterility.
- Enzymes Molecules which can act as catalysts in biological reactions.
- F 1 Hybrid The first generation progeny of the cross of two nonisogenic plants.
- Genotype The genetic constitution of a cell or organism.
- Haploid A cell or organism having one set of the two sets of chromosomes in a diploid.
- Linkage A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent.
- Marker A readily detectable phenotype, preferably inherited in codominant fashion (both alleles at a locus in a diploid heterozygote are readily detectable), with no environmental variance component, i.e., heritability of 1.
- Phenotype The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.
- Quantitative Trait Loci Quantitative trait loci (QTL) refer to genetic loci that control to some degree numerically representable traits that are usually continuously distributed.
- Resistance As used herein, the terms “resistance” and “tolerance” are used interchangeably to describe plants that show no symptoms to a specified biotic pest, pathogen, abiotic influence or environmental condition. These terms are also used to describe plants showing some symptoms but that are still able to produce marketable product with an acceptable yield. Some plants that are referred to as resistant or tolerant are only so in the sense that they may still produce a crop, even though the plants are stunted and the yield is reduced.
- Regeneration The development of a plant from tissue culture.
- RHS color chart value The RHS color chart is a standardized reference which allows accurate identification of any color. A color's designation on the chart describes its hue, brightness and saturation. A color is precisely named by the RHS color chart by identifying the group name, sheet number and letter, e.g., Yellow-Orange Group 19A or Red Group 41B.
- Self-pollination The transfer of pollen from the anther to the stigma of the same plant.
- Single Locus Converted (Conversion) Plant Plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a pepper variety are recovered in addition to the characteristics of the single locus transferred into the variety via the backcrossing technique and/or by genetic transformation.
- Tissue Culture A composition comprising isolated cells of the same or a different type or a collection of such cells organized into parts of a plant.
- Transgene A genetic locus comprising a sequence which has been introduced into the genome of a pepper plant by transformation.
- ATCC American Type Culture Collection
- the accession numbers for those deposited seeds of pepper hybrid SV3301HW and inbred parent line HHY-FD-1186 are ATCC Accession No. PTA-121391 and ATCC Accession No. PTA-121392, respectively.
- all restrictions upon the deposits will be removed, and the deposits are intended to meet all of the requirements of 37 C.F.R. ⁇ 1.801-1.809.
- the deposits will be maintained in the depository for a period of 30 years, or 5 years after the last request, or for the effective life of the patent, whichever is longer, and will be replaced if necessary during that period.
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Abstract
Description
- This application claims the priority of U.S. Provisional Appl. Ser. No. 62/045,464, filed Sep. 3, 2014, the entire disclosure of which is incorporated herein by reference.
- The present invention relates to the field of plant breeding and, more specifically, to the development of pepper hybrid SV3301HW and the inbred pepper lines SHY-114-1002 and HHY-FD-1186.
- The goal of vegetable breeding is to combine various desirable traits in a single variety/hybrid. Such desirable traits may include any trait deemed beneficial by a grower and/or consumer, including greater yield, resistance to insects or disease, tolerance to environmental stress, and nutritional value.
- Breeding techniques take advantage of a plant's method of pollination. There are two general methods of pollination: a plant self-pollinates if pollen from one flower is transferred to the same or another flower of the same plant or plant variety. A plant cross-pollinates if pollen comes to it from a flower of a different plant variety.
- Plants that have been self-pollinated and selected for type over many generations become homozygous at almost all gene loci and produce a uniform population of true breeding progeny, a homozygous plant. A cross between two such homozygous plants of different genotypes produces a uniform population of hybrid plants that are heterozygous for many gene loci. Conversely, a cross of two plants each heterozygous at a number of loci produces a population of hybrid plants that differ genetically and are not uniform. The resulting non-uniformity makes performance unpredictable.
- The development of uniform varieties requires the development of homozygous inbred plants, the crossing of these inbred plants, and the evaluation of the crosses. Pedigree breeding and recurrent selection are examples of breeding methods that have been used to develop inbred plants from breeding populations. Those breeding methods combine the genetic backgrounds from two or more plants or various other broad-based sources into breeding pools from which new lines and hybrids derived therefrom are developed by selfing and selection of desired phenotypes. The new lines and hybrids are evaluated to determine which of those have commercial potential.
- In one aspect, the present invention provides a pepper plant of the hybrid designated SV3301HW, the pepper line SHY-114-1002 or pepper line HHY-FD-1186. Also provided are pepper plants having all the physiological and morphological characteristics of such a plant. Parts of these pepper plants are also provided, for example, including pollen, an ovule, scion, a rootstock, a fruit, and a cell of the plant.
- In another aspect of the invention, a plant of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 comprising an added heritable trait is provided. The heritable trait may comprise a genetic locus that is, for example, a dominant or recessive allele. In one embodiment of the invention, a plant of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 is defined as comprising a single locus conversion. In specific embodiments of the invention, an added genetic locus confers one or more traits such as, for example, herbicide tolerance, insect resistance, disease resistance, and modified carbohydrate metabolism. In further embodiments, the trait may be conferred by a naturally occurring gene introduced into the genome of a line by backcrossing, a natural or induced mutation, or a transgene introduced through genetic transformation techniques into the plant or a progenitor of any previous generation thereof. When introduced through transformation, a genetic locus may comprise one or more genes integrated at a single chromosomal location.
- The invention also concerns the seed of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186. The pepper seed of the invention may be provided as an essentially homogeneous population of pepper seed of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186. Essentially homogeneous populations of seed are generally free from substantial numbers of other seed. Therefore, seed of hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 may be defined as forming at least about 97% of the total seed, including at least about 98%, 99% or more of the seed. The seed population may be separately grown to provide an essentially homogeneous population of pepper plants designated SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186.
- In yet another aspect of the invention, a tissue culture of regenerable cells of a pepper plant of hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 is provided. The tissue culture will preferably be capable of regenerating pepper plants capable of expressing all of the physiological and morphological characteristics of the starting plant, and of regenerating plants having substantially the same genotype as the starting plant. Examples of some of the physiological and morphological characteristics of the hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 include those traits set forth in the tables herein. The regenerable cells in such tissue cultures may be derived, for example, from embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistils, flowers, seed and stalks. Still further, the present invention provides pepper plants regenerated from a tissue culture of the invention, the plants having all the physiological and morphological characteristics of hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186.
- In still yet another aspect of the invention, processes are provided for producing pepper seeds, plants and fruit, which processes generally comprise crossing a first parent pepper plant with a second parent pepper plant, wherein at least one of the first or second parent pepper plants is a plant of pepper line SHY-114-1002 or pepper line HHY-FD-1186. These processes may be further exemplified as processes for preparing hybrid pepper seed or plants, wherein a first pepper plant is crossed with a second pepper plant of a different, distinct genotype to provide a hybrid that has, as one of its parents, a plant of pepper line SHY-114-1002 or pepper line HHY-FD-1186. In these processes, crossing will result in the production of seed. The seed production occurs regardless of whether the seed is collected or not.
- In one embodiment of the invention, the first step in “crossing” comprises planting seeds of a first and second parent pepper plant, often in proximity so that pollination will occur for example, mediated by insect vectors. Alternatively, pollen can be transferred manually. Where the plant is self-pollinated, pollination may occur without the need for direct human intervention other than plant cultivation.
- A second step may comprise cultivating or growing the seeds of first and second parent pepper plants into plants that bear flowers. A third step may comprise preventing self-pollination of the plants, such as by emasculating the flowers (i.e., killing or removing the pollen).
- A fourth step for a hybrid cross may comprise cross-pollination between the first and second parent pepper plants. Yet another step comprises harvesting the seeds from at least one of the parent pepper plants. The harvested seed can be grown to produce a pepper plant or hybrid pepper plant.
- The present invention also provides the pepper seeds and plants produced by a process that comprises crossing a first parent pepper plant with a second parent pepper plant, wherein at least one of the first or second parent pepper plants is a plant of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186. In one embodiment of the invention, pepper seed and plants produced by the process are first generation (F1) hybrid pepper seed and plants produced by crossing a plant in accordance with the invention with another, distinct plant. The present invention further contemplates plant parts of such an F1 hybrid pepper plant, and methods of use thereof. Therefore, certain exemplary embodiments of the invention provide an F1 hybrid pepper plant and seed thereof.
- In still yet another aspect, the present invention provides a method of producing a plant derived from hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186, the method comprising the steps of: (a) preparing a progeny plant derived from hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186, wherein said preparing comprises crossing a plant of the hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 with a second plant; and (b) crossing the progeny plant with itself or a second plant to produce a seed of a progeny plant of a subsequent generation. In further embodiments, the method may additionally comprise: (c) growing a progeny plant of a subsequent generation from said seed of a progeny plant of a subsequent generation and crossing the progeny plant of a subsequent generation with itself or a second plant; and repeating the steps for an additional 3-10 generations to produce a plant derived from hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186. The plant derived from hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 may be an inbred line, and the aforementioned repeated crossing steps may be defined as comprising sufficient inbreeding to produce the inbred line. In the method, it may be desirable to select particular plants resulting from step (c) for continued crossing according to steps (b) and (c). By selecting plants having one or more desirable traits, a plant derived from hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 is obtained which possesses some of the desirable traits of the line/hybrid as well as potentially other selected traits.
- In certain embodiments, the present invention provides a method of producing food or feed comprising: (a) obtaining a plant of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186, wherein the plant has been cultivated to maturity, and (b) collecting at least one pepper from the plant.
- In still yet another aspect of the invention, the genetic complement of pepper hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 is provided. The phrase “genetic complement” is used to refer to the aggregate of nucleotide sequences, the expression of which sequences defines the phenotype of, in the present case, a pepper plant, or a cell or tissue of that plant. A genetic complement thus represents the genetic makeup of a cell, tissue or plant, and a hybrid genetic complement represents the genetic make up of a hybrid cell, tissue or plant. The invention thus provides pepper plant cells that have a genetic complement in accordance with the pepper plant cells disclosed herein, and seeds and plants containing such cells.
- Plant genetic complements may be assessed by genetic marker profiles, and by the expression of phenotypic traits that are characteristic of the expression of the genetic complement, e.g., isozyme typing profiles. It is understood that hybrid SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 could be identified by any of the many well known techniques such as, for example, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).
- In still yet another aspect, the present invention provides hybrid genetic complements, as represented by pepper plant cells, tissues, plants, and seeds, formed by the combination of a haploid genetic complement of a pepper plant of the invention with a haploid genetic complement of a second pepper plant, preferably, another, distinct pepper plant. In another aspect, the present invention provides a pepper plant regenerated from a tissue culture that comprises a hybrid genetic complement of this invention.
- Any embodiment discussed herein with respect to one aspect of the invention applies to other aspects of the invention as well, unless specifically noted.
- The term “about” is used to indicate that a value includes the standard deviation of the mean for the device or method being employed to determine the value. The use of the term “or” in the claims is used to mean “and/or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive. When used in conjunction with the word “comprising” or other open language in the claims, the words “a” and “an” denote “one or more,” unless specifically noted otherwise. The terms “comprise,” “have” and “include” are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as “comprises,” “comprising,” “has,” “having,” “includes” and “including,” are also open-ended. For example, any method that “comprises,” “has” or “includes” one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps. Similarly, any plant that “comprises,” “has” or “includes” one or more traits is not limited to possessing only those one or more traits and covers other unlisted traits.
- Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and any specific examples provided, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.
- The invention provides methods and compositions relating to plants, seeds and derivatives of pepper hybrid SV3301HW, pepper line SHY-114-1002 and pepper line HHY-FD-1186.
- Hybrid SV3301HW is a hot Hungarian wax pepper with typical fruit type, fruit shape, smell, pungency, and flavor of a hot Hungarian wax. It has higher yield, 783 g/plant, compared to 713 g/plant for the standard hybrid Inferno. SV3301HW produces 17.3 fruits per plant while Inferno produces 15.3 fruits per plant. This hybrid is grown primarily in the U.S. for harvest as white fruits for processing and fresh market, with secondary use as a fresh pepper in Mexico. It exhibits excellent fresh and processed fruit quality due to its bright white color and moderate capsaicin content.
- The fruits of hybrid SV3301HW average 18.6 cm long×3.4 cm wide, with typical Hungarian wax shape and smell. Immature color is white (very pale yellow in some areas), and fruits mature to red. Fruit pungency of hybrid SV3301HW is moderate, around 640 Scoville Heat Units (fresh basis). Fruit weight of hybrid SV3301HW is 45.2 g while that of ‘Inferno’ is 46.7 g. Plant height averaged 70 cm, with a semi-spreading plant habit. It is ready to harvest at the white stage at approximately 75 days after transplanting, compared to 72 days for the standard ‘Inferno’. SV3301HW is well-adapted to the main hot pepper growing areas of the U.S. and Mexico, with excellent fresh and processed fruit quality. Hybrid SV3301HW is resistant to Tomato Spotted Wilt Virus (TSWV) and races 0-3 of Xanthomonas campestris vesacatoria (X3R), whereas Inferno is susceptible to both.
- The parents of hybrid SV3301HW are SHY-114-1002 and HHY-FD-1186. The parent lines are uniform and stable, as is a hybrid produced therefrom. A small percentage of variants can occur within commercially acceptable limits for almost any characteristic during the course of repeated multiplication. However no variants are expected.
- In accordance with one aspect of the present invention, there is provided a plant having the physiological and morphological characteristics of pepper hybrid SV3301HW and the parent lines thereof.
- A description of the physiological and morphological characteristics of pepper hybrid SV3301HW and pepper line HHY-FD-1186 is presented in Tables 1-2.
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TABLE 1 Physiological and Morphological Characteristics of Line HHY-FD-1186 CHARACTERISTIC HHY-FD-1186 HUNGARIAN 1. Species C. annuum C. annuum 2. Maturity (in region of best adaptability) days from transplanting until 48 45 mature green stage days from transplanting until 82 78 mature red or yellow stage days from direct seeding until 88 85 mature green stage days from direct seeding until 122 117 mature red or yellow stage 3. Plant habit semi-spreading semi-spreading attitude upright/erect semi-upright/semi- (De Cayenne, Doux très erect long des Landes, Piquant d'Algérie) plant height 60.6 cm 48.7 cm plant width 47.2 cm 52.4 cm length of stem from cotyledon 19.1 cm 11.7 cm to first flower length of the third internode 84.4 mm 69.6 mm (from soil surface) length of stem medium (Belsir, medium Lamuyo) shortened internode (in upper present present part) (Fehér, Kalocsai 601, Kalocsai 702) number of internodes between more than three one to three the first flower and the (Kalocsai 702) shortened internodes stem: hairiness of nodes medium (Doux très long absent or very weak des Landes, Farnese) height medium (HRF) short basal branches few (2-3) none branch flexibility rigid (Yolo Wonder) willowy stem strength (breakage weak intermediate resistance) 4. Leaf length of blade medium (Atol, Blondy, short Marconi, Merit, Anthea) width of blade medium (Albargia, narrow Balaton, Danubia, Marconi, Merit) width 64.3 mm 31.2 mm length 116.6 mm 108.7 mm petiole length 54.1 mm 39.0 mm color dark green dark green color (RHS Color Chart value) 147A 147A intensity of green color dark (Dolmy, Tinto) dark mature leaf shape ovate (Balico, Sonar) lanceolate leaf and stem pubescence absent light undulation of margin medium (Tenor) very weak blistering very weak very weak profile in cross section moderately concave strongly concave (Doux italien, Favolor) glossiness weak (De Cayenne, medium Doux très long des Favolor) 5. Flower peduncle: attitude semi-drooping (Blondy) erect flowers per leaf axil 1.2 1 calyx lobes 6.6 6.4 petals 6.9 6.3 diameter 29.9 mm 28.9 mm corolla color white white corolla throat markings yellow yellow anther color purple purple style length exceeds stamen exceeds stamen self-incompatibility present present 6. Fruit group Long Way (Sweet Short Wax (Floral Banana) Gem) color (before maturity) yellow (Fehér, Sweet yellow banana) intensity of color (before medium light maturity) immature fruit color medium green yellow immature fruit color (RHS 1-B 1-B Color Chart value) attitude/position drooping/pendent (De drooping/pendent Cayenne, Lamuyo) length long (Doux d'Espagne, medium Majister) diameter medium (Doux italien, narrow Corno di toro) ratio length/diameter medium (Adra, Cherry large Sweet, Daniel, Delphin, Edino) calyx diameter 26.7 mm 26.7 mm fruit length 173.7 mm 121.9 mm fruit diameter at calyx 36.1 mm 32.2 mm attachment fruit diameter at mid-point 30.9 mm 34.6 mm flesh thickness at mid-point 2.9 mm 3.1 mm average number of fruits per 20.7 29.8 plant % large fruits 29.50 (weight range: 13.60 (weight range: 40.5 to 80) 40.5 to 60) % medium fruits 42.40 (weight range: 20 62.10 (weight range: to 39.5) 20 to 39.5) % small fruits 28.10 (weight range: 1 24.30 (weight range: 1 to 19) to 19) average fruit weight 32.4 gm 27.8 gm fruit shape (longitudinal narrowly triangular (De narrowly triangular section) Cayenne, Demon) fruit shape (cross section, at angular/tri-angular circular level of placenta) sinuation of pericarp at basal medium (Duna, Banán) weak part sinuation of pericarp excluding absent or very weak weak basal part (Delphin, Milord) texture of surface slightly wrinkled smooth or slightly (Doux très long des wrinkled Landes) color (at maturity) red red (Fehér, Lamuyo) intensity of color (at maturity) dark dark mature fruit color red red mature fruit color (RHS Color 46A N64A Chart value) glossiness strong shiny (Doux italien, Trophy) stalk cavity absent (Corinto, Corno absent di toro, Sweet banana, Sucette de Provence) pedicel length 28.5 mm 29.0 mm pedicel thickness 5.8 mm 4.7 mm pedicel shape curved straight pedicel cavity absent absent stalk: length medium medium (Fehér, Sonar) stalk: thickness medium medium (Doux italien, Surpas) base shape rounded rounded shape of apex moderately acute moderately acute shape Elongate (Long Thin Elongate (Long Thin Cayenne) Cayenne) set concentrated concentrated depth of interloculary grooves shallow very shallow (Milord, Topgirl) number of locules predominantly three predominantly three (Century) % fruits with one locule 0% 0% % fruits with two locules 6.60% 20.00% % fruits with three locules 83% 67% % fruits with four locules 10.00% 13.30% % fruits with five locules 0% 0% average number of locules 3 2.9 thickness of flesh medium thin (Fehér, Lamuyo) calyx: aspect enveloping (cup- enveloping (cup- shaped) shaped) (De Cayenne, Sweet banana) pungency hot hot capsaicin in placenta present (De Cayenne) present mg Capsaicin per gram dry fruit 0.65 mg 0.25 mg Scoville Units (dry fruit) 10,185 3,987 flavor strong pepper flavor strong pepper flavor glossiness shiny shiny 7. Seed seed cavity length 154.8 mm 107.7 mm seed cavity diameter 26.9 mm 27.8 mm placenta length 114.2 mm 76.9 mm number of seeds per fruit 108.8 116.3 grams per 1000 seeds 7.4 gm 6.4 gm color yellow yellow 8. Anthocyanin coloration of: Seedling: anthocyanin absent weak coloration of hypocotyl (Albaregia, Albena) Plant: anthocyanin coloration of strong moderate stem Plant: anthocyanin coloration of strong (California strong nodes wonder) stem: intensity of anthocyanin strong strong coloration of nodes (Piquant d′Algérie, Zarai) leaf weak moderate pedicel absent weak calyx weak absent anther present (Lamuyo) present fruit coloration absent (Lamuyo) absent beginning of flowering (1st medium early flower on 2nd flowering node) (Lamuyo, Latino) time of maturity early early (Fehér, Lady Bell, Topgirl) *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention. -
TABLE 2 Physiological and Morphological Characteristics of Pepper Hybrid SV3301HW Sweet Banana (SZZ CHARACTERISTIC SV3301HW 35-1051) 1. Species C. annuum annuum 2. Maturity (in region of best adaptability) days from transplanting until 46 46 mature green stage days from transplanting until 88 83 mature red or yellow stage days from direct seeding until 89 88 mature green stage days from direct seeding until 121 126 mature red or yellow stage 3. Plant habit compact compact attitude upright/erect upright/erect plant height 65.9 cm 64.9 cm plant width 60.5 cm 52.3 cm length of stem from cotyledon 15.0 cm 21.1 cm to first flower length of the third internode 80.0 mm 118.0 mm (from soil surface) length of stem medium long shortened internode (in upper present absent part) number of internodes between one to three the first flower and the shortened internodes stem: hairiness of nodes weak weak height tall tall basal branches few (2-3) none branch flexibility willowy rigid stem strength (breakage intermediate weak resistance) 4. Leaf length of blade short medium width of blade medium medium width 61.4 mm 61.6 mm length 111.3 mm 119.2 mm petiole length 48.7 mm 55.8 mm color dark green dark green color (RHS Color Chart value) 137A 137A intensity of green color dark dark mature leaf shape ovate lanceolate leaf and stem pubescence light absent undulation of margin medium medium blistering medium medium profile in cross section strongly concave moderately concave glossiness strong medium 5. Flower peduncle: attitude semi-drooping semi-drooping flowers per leaf axil 1 1 calyx lobes 5.5 5.9 petals 5.6 6 diameter 20.2 mm 22.4 mm corolla color white white fruit diameter at calyx 33.2 mm 36.1 mm attachment fruit diameter at mid-point 32.2 mm 30.7 mm flesh thickness at mid-point 2.7 mm 2.9 mm average number of fruits per 22.7 25.3 plant % large fruits 30.50% weight range 50 4.50% weight range to 90 50 to 58 % medium fruits 39.90% weight range 30 38.40% weight range to 49 30 to 49 % small fruits 29.60% weight range 1 57.10% weight range to 29 1 to 29 average fruit weight 64.8 gm 41.6 gm fruit shape (longitudinal narrowly triangular horn-shaped section) fruit shape (cross section, at angular/triangular angular/triangular level of placenta) sinuation of pericarp at basal weak absent or very weak part sinuation of pericarp excluding weak absent or very weak basal part texture of surface slightly wrinkled smooth or very slightly wrinkled surface smoothness rough rough color (at maturity) red red intensity of color (at maturity) dark dark mature fruit color red red mature fruit color (RHS Color 46A 46A Chart value) glossiness strong strong stalk cavity absent absent pedicel length 48.3 mm 36.2 mm pedicel thickness 5.6 mm 5.3 mm pedicel shape curved curved pedicel cavity absent absent stalk: length long medium stalk: thickness medium medium base shape rounded rounded apex shape blunt blunt shape of apex very acute very acute/pointed shape Elongate (Long Thin Elongate (Long Thin Cayenne) Cayenne) set concentrated concentrated depth of interloculary grooves shallow medium number of locules predominantly three predominantly four and more % fruits with one locule 0% 0% % fruits with two locules 0% 0% % fruits with three locules 73.30% 40% % fruits with four locules 26.70% 53% % fruits with five or more 0% 6.70% locules average number of locules 3.3 3.6 thickness of flesh thin thin calyx: aspect enveloping (cup- enveloping (cup- shaped) shaped) pungency hot hot capsaicin in placenta present present flavor strong pepper flavor strong pepper flavor glossiness shiny shiny 7. Seed seed cavity length 172.6 mm 118.4 mm seed cavity diameter 24.4 mm 22.0 mm placenta length 88.3 mm 95.9 mm number of seeds per fruit 216.9 254.9 grams per 1000 seeds 6.0 gm 6.0 gm color yellow yellow 8. Anthocyanin coloration of: hypocotyl absent moderate stem absent absent node moderate moderate stem: intensity of anthocyanin medium strong coloration of nodes leaf moderate absent pedicel weak weak calyx absent absent anther present present fruit coloration absent weak beginning of flowering (1st medium medium flower on 2nd flowering node) time of maturity early early *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention. - One aspect of the current invention concerns methods for producing seed of pepper hybrid SV3301HW involving crossing pepper lines SHY-114-1002 and HHY-FD-1186. Alternatively, in other embodiments of the invention, hybrid SV3301HW, line SHY-114-1002, or line HHY-FD-1186 may be crossed with itself or with any second plant. Such methods can be used for propagation of hybrid SV3301HW and/or the pepper lines SHY-114-1002 and HHY-FD-1186, or can be used to produce plants that are derived from hybrid SV3301HW and/or the pepper lines SHY-114-1002 and HHY-FD-1186. Plants derived from hybrid SV3301HW and/or the pepper lines SHY-114-1002 and HHY-FD-1186 may be used, in certain embodiments, for the development of new pepper varieties.
- The development of new varieties using one or more starting varieties is well known in the art. In accordance with the invention, novel varieties may be created by crossing hybrid SV3301HW followed by multiple generations of breeding according to such well known methods. New varieties may be created by crossing with any second plant. In selecting such a second plant to cross for the purpose of developing novel lines, it may be desired to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Once initial crosses have been made, inbreeding and selection take place to produce new varieties. For development of a uniform line, often five or more generations of selfing and selection are involved.
- Uniform lines of new varieties may also be developed by way of double-haploids. This technique allows the creation of true breeding lines without the need for multiple generations of selfing and selection. In this manner true breeding lines can be produced in as little as one generation. Haploid embryos may be produced from microspores, pollen, anther cultures, or ovary cultures. The haploid embryos may then be doubled autonomously, or by chemical treatments (e.g. colchicine treatment). Alternatively, haploid embryos may be grown into haploid plants and treated to induce chromosome doubling. In either case, fertile homozygous plants are obtained. In accordance with the invention, any of such techniques may be used in connection with a plant of the invention and progeny thereof to achieve a homozygous line.
- Backcrossing can also be used to improve an inbred plant. Backcrossing transfers a specific desirable trait from one inbred or non-inbred source to an inbred that lacks that trait. This can be accomplished, for example, by first crossing a superior inbred (A) (recurrent parent) to a donor inbred (non-recurrent parent), which carries the appropriate locus or loci for the trait in question. The progeny of this cross are then mated back to the superior recurrent parent (A) followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent. After five or more backcross generations with selection for the desired trait, the progeny have the characteristic being transferred, but are like the superior parent for most or almost all other loci. The last backcross generation would be selfed to give pure breeding progeny for the trait being transferred.
- The plants of the present invention are particularly well suited for the development of new lines based on the elite nature of the genetic background of the plants. In selecting a second plant to cross with SV3301HW and/or pepper lines SHY-114-1002 and HHY-FD-1186 for the purpose of developing novel pepper lines, it will typically be preferred to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Examples of desirable traits may include, in specific embodiments, high seed yield, high seed germination, seedling vigor, high fruit yield, disease tolerance or resistance, and adaptability for soil and climate conditions. Consumer-driven traits, such as a fruit shape, color, texture, and taste are other examples of traits that may be incorporated into new lines of pepper plants developed by this invention.
- As described above, hybrid SV3301HW exhibits desirable traits, as conferred by pepper lines SHY-114-1002 and HHY-FD-1186. The performance characteristics of hybrid SV3301HW and pepper lines SHY-114-1002 and HHY-FD-1186 were the subject of an objective analysis of the performance traits relative to other varieties. The results of the analysis are presented in Table 2.
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TABLE 2 Performance data for Hybrid SV3301HW and comparative varieties. Color Matu- imma- rity Yld Len Wid Av. wt % dry Variety ture (DAT) g/plt cm cm g/fruit matter Inferno White 72 713 17.9 3.6 46.7 6.9 Hot Banana White 78 484 17.4 3.4 40.2 7.0 SV3301HW White 75 783 18.6 3.4 45.2 7.0 - In certain aspects of the invention, plants described herein are provided modified to include at least a first desired heritable trait. Such plants may, in one embodiment, be developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to a genetic locus transferred into the plant via the backcrossing technique. The term single locus converted plant as used herein refers to those pepper plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to the single locus transferred into the variety via the backcrossing technique. By essentially all of the morphological and physiological characteristics, it is meant that the characteristics of a plant are recovered that are otherwise present when compared in the same environment, other than an occasional variant trait that might arise during backcrossing or direct introduction of a transgene.
- Backcrossing methods can be used with the present invention to improve or introduce a characteristic into the present variety. The parental pepper plant which contributes the locus for the desired characteristic is termed the nonrecurrent or donor parent. This terminology refers to the fact that the nonrecurrent parent is used one time in the backcross protocol and therefore does not recur. The parental pepper plant to which the locus or loci from the nonrecurrent parent are transferred is known as the recurrent parent as it is used for several rounds in the backcrossing protocol.
- In a typical backcross protocol, the original variety of interest (recurrent parent) is crossed to a second variety (nonrecurrent parent) that carries the single locus of interest to be transferred. The resulting progeny from this cross are then crossed again to the recurrent parent and the process is repeated until a pepper plant is obtained wherein essentially all of the morphological and physiological characteristics of the recurrent parent are recovered in the converted plant, in addition to the single transferred locus from the nonrecurrent parent.
- The selection of a suitable recurrent parent is an important step for a successful backcrossing procedure. The goal of a backcross protocol is to alter or substitute a single trait or characteristic in the original variety. To accomplish this, a single locus of the recurrent variety is modified or substituted with the desired locus from the nonrecurrent parent, while retaining essentially all of the rest of the desired genetic, and therefore the desired physiological and morphological constitution of the original variety. The choice of the particular nonrecurrent parent will depend on the purpose of the backcross; one of the major purposes is to add some commercially desirable trait to the plant. The exact backcrossing protocol will depend on the characteristic or trait being altered and the genetic distance between the recurrent and nonrecurrent parents. Although backcrossing methods are simplified when the characteristic being transferred is a dominant allele, a recessive allele, or an additive allele (between recessive and dominant), may also be transferred. In this instance it may be necessary to introduce a test of the progeny to determine if the desired characteristic has been successfully transferred.
- In one embodiment, progeny pepper plants of a backcross in which a plant described herein is the recurrent parent comprise (i) the desired trait from the non-recurrent parent and (ii) all of the physiological and morphological characteristics of pepper the recurrent parent as determined at the 5% significance level when grown in the same environmental conditions.
- New varieties can also be developed from more than two parents. The technique, known as modified backcrossing, uses different recurrent parents during the backcrossing. Modified backcrossing may be used to replace the original recurrent parent with a variety having certain more desirable characteristics or multiple parents may be used to obtain different desirable characteristics from each.
- With the development of molecular markers associated with particular traits, it is possible to add additional traits into an established germ line, such as represented here, with the end result being substantially the same base germplasm with the addition of a new trait or traits. Molecular breeding, as described in Moose and Mumm, 2008 (Plant Physiology, 147: 969-977), for example, and elsewhere, provides a mechanism for integrating single or multiple traits or QTL into an elite line. This molecular breeding-facilitated movement of a trait or traits into an elite line may encompass incorporation of a particular genomic fragment associated with a particular trait of interest into the elite line by the mechanism of identification of the integrated genomic fragment with the use of flanking or associated marker assays. In the embodiment represented here, one, two, three or four genomic loci, for example, may be integrated into an elite line via this methodology. When this elite line containing the additional loci is further crossed with another parental elite line to produce hybrid offspring, it is possible to then incorporate at least eight separate additional loci into the hybrid. These additional loci may confer, for example, such traits as a disease resistance or a fruit quality trait. In one embodiment, each locus may confer a separate trait. In another embodiment, loci may need to be homozygous and exist in each parent line to confer a trait in the hybrid. In yet another embodiment, multiple loci may be combined to confer a single robust phenotype of a desired trait.
- Many single locus traits have been identified that are not regularly selected for in the development of a new inbred but that can be improved by backcrossing techniques. Single locus traits may or may not be transgenic; examples of these traits include, but are not limited to, herbicide resistance, resistance to bacterial, fungal, or viral disease, insect resistance, modified fatty acid or carbohydrate metabolism, and altered nutritional quality. These comprise genes generally inherited through the nucleus.
- Direct selection may be applied where the single locus acts as a dominant trait. For this selection process, the progeny of the initial cross are assayed for viral resistance and/or the presence of the corresponding gene prior to the backcrossing. Selection eliminates any plants that do not have the desired gene and resistance trait, and only those plants that have the trait are used in the subsequent backcross. This process is then repeated for all additional backcross generations.
- Selection of pepper plants for breeding is not necessarily dependent on the phenotype of a plant and instead can be based on genetic investigations. For example, one can utilize a suitable genetic marker which is closely genetically linked to a trait of interest. One of these markers can be used to identify the presence or absence of a trait in the offspring of a particular cross, and can be used in selection of progeny for continued breeding. This technique is commonly referred to as marker assisted selection. Any other type of genetic marker or other assay which is able to identify the relative presence or absence of a trait of interest in a plant can also be useful for breeding purposes. Procedures for marker assisted selection are well known in the art. Such methods will be of particular utility in the case of recessive traits and variable phenotypes, or where conventional assays may be more expensive, time consuming or otherwise disadvantageous. Types of genetic markers which could be used in accordance with the invention include, but are not necessarily limited to, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).
- Many useful traits that can be introduced by backcrossing, as well as directly into a plant, are those which are introduced by genetic transformation techniques. Genetic transformation may therefore be used to insert a selected transgene into a plant of the invention or may, alternatively, be used for the preparation of transgenes which can be introduced by backcrossing. Methods for the transformation of plants that are well known to those of skill in the art and applicable to many crop species include, but are not limited to, electroporation, microprojectile bombardment, Agrobacterium-mediated transformation and direct DNA uptake by protoplasts.
- To effect transformation by electroporation, one may employ either friable tissues, such as a suspension culture of cells or embryogenic callus or alternatively one may transform immature embryos or other organized tissue directly. In this technique, one would partially degrade the cell walls of the chosen cells by exposing them to pectin-degrading enzymes (pectolyases) or mechanically wound tissues in a controlled manner.
- An efficient method for delivering transforming DNA segments to plant cells is microprojectile bombardment. In this method, particles are coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, platinum, and preferably, gold. For the bombardment, cells in suspension are concentrated on filters or solid culture medium. Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the macroprojectile stopping plate.
- An illustrative embodiment of a method for delivering DNA into plant cells by acceleration is the Biolistics Particle Delivery System, which can be used to propel particles coated with DNA or cells through a screen, such as a stainless steel or Nytex screen, onto a surface covered with target cells. The screen disperses the particles so that they are not delivered to the recipient cells in large aggregates. Microprojectile bombardment techniques are widely applicable, and may be used to transform virtually any plant species.
- Agrobacterium-mediated transfer is another widely applicable system for introducing gene loci into plant cells. An advantage of the technique is that DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations (Klee et al., Bio-Technology, 3(7):637-642, 1985). Moreover, recent technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate the construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes. Additionally, Agrobacterium containing both armed and disarmed Ti genes can be used for transformation.
- In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene locus transfer. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art (Fraley et al., Bio/Technology, 3:629-635, 1985; U.S. Pat. No. 5,563,055).
- Transformation of plant protoplasts also can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation, and combinations of these treatments (see, e.g., Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al., Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature, 312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986; Marcotte et al., Nature, 335:454, 1988). Transformation of plants and expression of foreign genetic elements is exemplified in Choi et al. (Plant Cell Rep., 13: 344-348, 1994), and Ellul et al. (Theor. Appl. Genet., 107:462-469, 2003).
- A number of promoters have utility for plant gene expression for any gene of interest including but not limited to selectable markers, scoreable markers, genes for pest tolerance, disease resistance, nutritional enhancements and any other gene of agronomic interest. Examples of constitutive promoters useful for plant gene expression include, but are not limited to, the cauliflower mosaic virus (CaMV) P-35S promoter, which confers constitutive, high-level expression in most plant tissues (see, e.g., Odel et al., Nature, 313:810, 1985), including in monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591, 1990; Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemly duplicated version of the CaMV 35S promoter, the enhanced 35S promoter (P-e35S);1 the nopaline synthase promoter (An et al., Plant Physiol., 88:547, 1988); the octopine synthase promoter (Fromm et al., Plant Cell, 1:977, 1989); and the figwort mosaic virus (P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter (P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem; the cauliflower mosaic virus 19S promoter; a sugarcane bacilliform virus promoter; a commelina yellow mottle virus promoter; and other plant DNA virus promoters known to express in plant cells.
- A variety of plant gene promoters that are regulated in response to environmental, hormonal, chemical, and/or developmental signals can also be used for expression of an operably linked gene in plant cells, including promoters regulated by (1) heat (Callis et al., Plant Physiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter, Schaffner and Sheen, Plant Cell, 3:997, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., EMBO J., 4:2723, 1985), (3) hormones, such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989), (4) wounding (e.g., wunl, Siebertz et al., Plant Cell, 1:961, 1989); or (5) chemicals such as methyl jasmonate, salicylic acid, or Safener. It may also be advantageous to employ organ-specific promoters (e.g., Roshal et al., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988; Bustos et al., Plant Cell, 1:839, 1989).
- Exemplary nucleic acids which may be introduced to plants of this invention include, for example, DNA sequences or genes from another species, or even genes or sequences which originate with or are present in the same species, but are incorporated into recipient cells by genetic engineering methods rather than classical reproduction or breeding techniques. However, the term “exogenous” is also intended to refer to genes that are not normally present in the cell being transformed, or perhaps simply not present in the form, structure, etc., as found in the transforming DNA segment or gene, or genes which are normally present and that one desires to express in a manner that differs from the natural expression pattern, e.g., to over-express. Thus, the term “exogenous” gene or DNA is intended to refer to any gene or DNA segment that is introduced into a recipient cell, regardless of whether a similar gene may already be present in such a cell. The type of DNA included in the exogenous DNA can include DNA which is already present in the plant cell, DNA from another plant, DNA from a different organism, or a DNA generated externally, such as a DNA sequence containing an antisense message of a gene, or a DNA sequence encoding a synthetic or modified version of a gene.
- Many hundreds if not thousands of different genes are known and could potentially be introduced into a pepper plant according to the invention. Non-limiting examples of particular genes and corresponding phenotypes one may choose to introduce into a pepper plant include one or more genes for insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pest tolerance such as genes for fungal disease control, herbicide tolerance such as genes conferring glyphosate tolerance, and genes for quality improvements such as yield, nutritional enhancements, environmental or stress tolerances, or any desirable changes in plant physiology, growth, development, morphology or plant product(s). For example, structural genes would include any gene that confers insect tolerance including but not limited to a Bacillus insect control protein gene as described in WO 99/31248, herein incorporated by reference in its entirety, U.S. Pat. No. 5,689,052, herein incorporated by reference in its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by reference in their entirety. In another embodiment, the structural gene can confer tolerance to the herbicide glyphosate as conferred by genes including, but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, herein incorporated by reference in its entirety, or glyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporated by reference in its entirety.
- Alternatively, the DNA coding sequences can affect these phenotypes by encoding a non-translatable RNA molecule that causes the targeted inhibition of expression of an endogenous gene, for example via antisense- or cosuppression-mediated mechanisms (see, for example, Bird et al., Biotech. Gen. Engin. Rev., 9:207, 1991). The RNA could also be a catalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desired endogenous mRNA product (see for example, Gibson and Shillito, Mol. Biotech., 7:125, 1997). Thus, any gene which produces a protein or mRNA which expresses a phenotype or morphology change of interest is useful for the practice of the present invention.
- In the description and tables herein, a number of terms are used. In order to provide a clear and consistent understanding of the specification and claims, the following definitions are provided:
- Allele: Any of one or more alternative forms of a gene locus, all of which alleles relate to one trait or characteristic. In a diploid cell or organism, the two alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes.
- Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (F1), back to one of the parents of the hybrid progeny. Backcrossing can be used to introduce one or more single locus conversions from one genetic background into another.
- Crossing: The mating of two parent plants.
- Cross-pollination: Fertilization by the union of two gametes from different plants.
- Diploid: A cell or organism having two sets of chromosomes.
- Emasculate: The removal of plant male sex organs or the inactivation of the organs with a cytoplasmic or nuclear genetic factor or a chemical agent conferring male sterility.
- Enzymes: Molecules which can act as catalysts in biological reactions.
- F1 Hybrid: The first generation progeny of the cross of two nonisogenic plants.
- Genotype: The genetic constitution of a cell or organism.
- Haploid: A cell or organism having one set of the two sets of chromosomes in a diploid.
- Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent.
- Marker: A readily detectable phenotype, preferably inherited in codominant fashion (both alleles at a locus in a diploid heterozygote are readily detectable), with no environmental variance component, i.e., heritability of 1.
- Phenotype: The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.
- Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer to genetic loci that control to some degree numerically representable traits that are usually continuously distributed.
- Resistance: As used herein, the terms “resistance” and “tolerance” are used interchangeably to describe plants that show no symptoms to a specified biotic pest, pathogen, abiotic influence or environmental condition. These terms are also used to describe plants showing some symptoms but that are still able to produce marketable product with an acceptable yield. Some plants that are referred to as resistant or tolerant are only so in the sense that they may still produce a crop, even though the plants are stunted and the yield is reduced.
- Regeneration: The development of a plant from tissue culture.
- Royal Horticultural Society (RHS) color chart value: The RHS color chart is a standardized reference which allows accurate identification of any color. A color's designation on the chart describes its hue, brightness and saturation. A color is precisely named by the RHS color chart by identifying the group name, sheet number and letter, e.g., Yellow-Orange Group 19A or Red Group 41B.
- Self-pollination: The transfer of pollen from the anther to the stigma of the same plant.
- Single Locus Converted (Conversion) Plant: Plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a pepper variety are recovered in addition to the characteristics of the single locus transferred into the variety via the backcrossing technique and/or by genetic transformation.
- Substantially Equivalent: A characteristic that, when compared, does not show a statistically significant difference (e.g., p=0.05) from the mean.
- Tissue Culture: A composition comprising isolated cells of the same or a different type or a collection of such cells organized into parts of a plant.
- Transgene: A genetic locus comprising a sequence which has been introduced into the genome of a pepper plant by transformation.
- A deposit of pepper hybrid SV3301HW and inbred parent line and HHY-FD-1186, disclosed above and recited in the claims, has been made with the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The dates of deposit was Jul. 16, 2014. The accession numbers for those deposited seeds of pepper hybrid SV3301HW and inbred parent line HHY-FD-1186 are ATCC Accession No. PTA-121391 and ATCC Accession No. PTA-121392, respectively. Upon issuance of a patent, all restrictions upon the deposits will be removed, and the deposits are intended to meet all of the requirements of 37 C.F.R. §1.801-1.809. The deposits will be maintained in the depository for a period of 30 years, or 5 years after the last request, or for the effective life of the patent, whichever is longer, and will be replaced if necessary during that period.
- Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the invention, as limited only by the scope of the appended claims.
- All references cited herein are hereby expressly incorporated herein by reference.
Claims (33)
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Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
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US9320214B2 (en) | 2013-11-21 | 2016-04-26 | Seminis Vegetable Seeds, Inc. | Pepper hybrid PX 11404796 |
CN105815143A (en) * | 2016-04-06 | 2016-08-03 | 牛福永 | Seedling-raising matrix for cultivating strong pepper seedlings |
CN105815144A (en) * | 2016-04-06 | 2016-08-03 | 牛福永 | Strong pepper seedling raising method |
US9532522B2 (en) | 2014-12-18 | 2017-01-03 | Seminis Vegetable Seeds, Inc. | Pepper hybrid SV7685PP |
US9699979B2 (en) | 2014-09-03 | 2017-07-11 | Seminis Vegetable Seeds, Inc. | Pepper hybrid PX-11459057 |
US9854758B2 (en) | 2016-01-06 | 2018-01-02 | Seminis Vegetable Seeds, Inc. | Pepper hybrid SV7073HT |
US9867343B2 (en) | 2016-02-24 | 2018-01-16 | Seminis Vegetable Seeds, Inc. | Pepper hybrid SV6496HJ |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
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US9907242B2 (en) | 2016-02-18 | 2018-03-06 | Seminis Vegetable Seeds, Inc. | Pepper hybrid SV5873HE |
US10058046B2 (en) | 2016-09-01 | 2018-08-28 | Seminis Vegetable Seeds, Inc. | Pepper hybrid SV3140HJ |
US10959392B2 (en) * | 2019-05-20 | 2021-03-30 | Seminis Vegetable Seeds, Inc. | Pepper hybrid SVPP8114 and parents thereof |
Citations (1)
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US20130024963A1 (en) * | 2011-07-22 | 2013-01-24 | Seminis Vegetable Seeds, Inc. | Pepper hybrid ps11447353 |
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Publication number | Priority date | Publication date | Assignee | Title |
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US8049078B2 (en) | 2007-08-30 | 2011-11-01 | Seminis Vegetable Seeds, Inc. | Habanero pepper hybrid PX11423487 |
US8071862B2 (en) | 2007-08-30 | 2011-12-06 | Seminis Vegetable Seeds, Inc. | Habanero pepper hybrid PX11423486 |
US8524991B2 (en) | 2009-07-30 | 2013-09-03 | Seminis Vegetable Seeds, Inc. | Pepper hybrid PX11435810 |
US8426702B2 (en) | 2010-08-16 | 2013-04-23 | Seminis Vegetable Seeds, Inc. | Pepper hybrid PS 11446271 |
US8415537B2 (en) | 2010-09-14 | 2013-04-09 | Seminis Vegetable Seeds, Inc. | Pepper hybrid PS11423700 |
US8835723B2 (en) | 2011-07-11 | 2014-09-16 | Seminis Vegetable Seeds, Inc. | Pepper hybrid PS11423123 |
US8962943B2 (en) | 2012-02-02 | 2015-02-24 | Seminis Vegetable Seeds, Inc. | Pepper hybrid PX11423782 |
US9320214B2 (en) | 2013-11-21 | 2016-04-26 | Seminis Vegetable Seeds, Inc. | Pepper hybrid PX 11404796 |
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US20130024963A1 (en) * | 2011-07-22 | 2013-01-24 | Seminis Vegetable Seeds, Inc. | Pepper hybrid ps11447353 |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9320214B2 (en) | 2013-11-21 | 2016-04-26 | Seminis Vegetable Seeds, Inc. | Pepper hybrid PX 11404796 |
US9699979B2 (en) | 2014-09-03 | 2017-07-11 | Seminis Vegetable Seeds, Inc. | Pepper hybrid PX-11459057 |
US9532522B2 (en) | 2014-12-18 | 2017-01-03 | Seminis Vegetable Seeds, Inc. | Pepper hybrid SV7685PP |
US9854758B2 (en) | 2016-01-06 | 2018-01-02 | Seminis Vegetable Seeds, Inc. | Pepper hybrid SV7073HT |
US9867343B2 (en) | 2016-02-24 | 2018-01-16 | Seminis Vegetable Seeds, Inc. | Pepper hybrid SV6496HJ |
CN105815143A (en) * | 2016-04-06 | 2016-08-03 | 牛福永 | Seedling-raising matrix for cultivating strong pepper seedlings |
CN105815144A (en) * | 2016-04-06 | 2016-08-03 | 牛福永 | Strong pepper seedling raising method |
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